The dissipation of fipronil, chlorpyrifos, fosthiazate and ethoprophos in soils from potato monoculture areas: first evidence for the enhanced biodegradation of fosthiazate.

BACKGROUND: A limited number of pesticides are available for the control of soil pests in potato. This, together with the monoculture nature of potato cultivation, does not favour chemical rotation, increasing the risk of reduced biological efficacy due to microbial adaptation. The dissipation of three major organophosphates (chlorpyrifos, ethoprophos and fosthiazate) was studied in comparison with fipronil, an insecticide recently introduced in potato cultivation, in 17 soils from potato monoculture areas in Greece to explore the extent of enhanced biodegradation development. RESULTS: The dissipation time of the four pesticides varied in the different soils, with DT50 values of 1.7-30.8 days, 2.7-56 days, 7.0-31.0 days and 24.5-116.5 days for fosthiazate, chlorpyrifos, ethoprophos and fipronil, respectively. A rapid dissipation of ethoprophos and fosthiazate in two soils with previous exposure to these nematicides provided first evidence for the development of enhanced biodegradation. Sterilisation of the given soils inhibited the dissipation of fosthiazate. Additionally, fosthiazate dissipation in the soils increased upon repeated applications. CONCLUSION: The development of enhanced biodegradation of fosthiazate in soils from potato monoculture regions was verified. This is the first report of enhanced biodegradation for this chemical. Further studies will focus on the isolation of microorganisms responsible for the dissipation of fosthiazate.

Fate of 14C-ethion insecticide in the presence of deltamethrin and dimilin pesticides in cotton seeds and oils, removal of ethion residues in oils, and bioavailability of its bound residues to experimental animals.

Ethyl-1-(14)C-ethion and some of its degradation products have been prepared for comparison purposes. Cotton plants were treated with (14)C-ethion alone and in the presence of deltamethrin and dimilin pesticides under conditions simulating local agricultural practice. (14)C-Residues in seeds were determined at harvest time; about 47.5% of (14)C-activity was associated with oil. After further extraction of seeds with ethanol, the ethanol-soluble (14)C-residues accounted for 10.6% of the total seed residues, whereas the cake contained about 37.3% of the total residues as bound residues in the case of ethion only. The bound residues decreased in the presence of deltamethrin and dimilin pesticides and amounted to 8.1 and 10.4% of the total residues, respectively. About 95% of the (14)C-activity in the crude oil could be eliminated by simulated commercial processes locally used for oil refining. Chromatographic analysis of crude cotton oil revealed the presence of ethion monooxon, O,O-diethyl phosphorothioate, and O,O-diethyl phosphoric acid in addition to one unknown compound in the case of ethion alone or ethion and dimilin. The same degradation products are found in the case of ethion and deltamethrin in addition to ethion dioxon, whereas ethanol extract revealed the presence of ethion dioxon and O,O-diethyl phosphoric acid as free metabolites. Acid hydrolysis of the conjugated metabolites in the ethanol extract yielded O,O-diethyl S-hydroxymethyl phosphorodithioate. The bound residues were quite readily bioavailable to the rats. After feeding rats with the cake containing ethion-bound residues, a substantial amount (60%) of (14)C-residues was eliminated in the urine, whereas the (14)C-residues excreted in expired air and feces were 10 and 9%, respectively. About 11% of the radioactive residues were distributed among various organs.

Identification and characterization of novel catalytic bioscavengers of organophosphorus nerve agents.

In an effort to discover novel catalytic bioscavengers of organophosphorus (OP) nerve agents, cell lysates from a diverse set of bacterial strains were screened for their capacity to hydrolyze the OP nerve agents VX, VR, and soman (GD). The library of bacterial strains was identified using both random and rational approaches. Specifically, two representative strains from eight categories of extremophiles were chosen at random. For the rational approach, the protein sequence of organophosphorus hydrolase (OPH) from Brevundimonas diminuta was searched against a non-redundant protein database using the Basic Local Alignment Search Tool to find regions of local similarity between sequences. Over 15 protein sequences with significant sequence similarity to OPH were identified from a variety of bacterial strains. Some of these matches were based on predicted protein structures derived from bacterial genome sequences rather than from bona fide proteins isolated from bacteria. Of the 25 strains selected for nerve agent testing, three bacterial strains had measurable levels of OP hydrolase activity. These strains are Ammoniphilus oxalaticus, Haloarcula sp., and Micromonospora aurantiaca. Lysates from A. oxalaticus had detectable hydrolysis of VR; Haloarcula sp. had appreciable hydrolysis of VX and VR, whereas lysates from M. aurantiaca had detectable hydrolysis of VR and GD.

Pepper leaf matrix as a promising analyte protectant prior to the analysis of thermolabile terbufos and its metabolites in pepper using GC-FPD.

During gas chromatography (GC), the matrix can deactivate the active site during the transport of the compound from the injector to the detector. This deactivation capacity varies among matrices, as it is dependant on the concentrations of the different constituent compounds of each matrix. During the analysis of terbufos and its metabolites, two of its metabolites were highly thermolabile, and were readily decomposed inside the GC system. As the matrix can mask the active site, we carried out a matrix-matched calibration in an effort to protect the analyte against decomposition. As a component of our analysis, the pepper matrix was the first to be matched; however, it failed to completely protect the metabolites. Subsequently, a variety of different compounds, including 3-ethoxy-1,2-propanediol, gulonolactone, and sorbitol at 10, 1, and 1mg/mL were tested; however, none of these generated the desired effect. We surmised that some of the compounds may have decomposed inside the injection port, so we introduced a carbofrit inlet liner, which is highly inert. But, this step did not improve the protective qualities of the matrices. Finally, pepper leaf matrix was added to the pepper matrix, and we observed a profound protective effect for almost all of the analytes tested. A selective detector (flame photometric detector with phosphorus filter) was used to facilitate a high matrix concentration without interaction with the analyte. After resolving the problem of these two metabolites, terbufos and its five toxic metabolites were analyzed in pepper and pepper leaf samples. The recovery rates for terbufos and its metabolites were 73-114.5% with a relative standard deviation of <12%. This method was successfully applied to field samples, and terbufos sulfone, terbufos sulfoxide, and terbufoxon sulfoxide were found as residues in the suspected pepper and pepper leaf samples.

Non-specific biodegradation of the organophosphorus pesticides, cadusafos and ethoprophos, by two bacterial isolates.

An enrichment culture technique was used for the isolation of microorganisms responsible for the enhanced biodegradation of the nematicide cadusafos in soils from a potato monoculture area in Northern Greece. Mineral salts medium supplemented with nitrogen (MSMN), where cadusafos (10 mg l(-1)) was the sole carbon source, and soil extract medium (SEM) were used for the isolation of cadusafos-degrading bacteria. Two pure bacterial cultures, named CadI and CadII, were isolated and subsequently characterized by sequencing of 16S rRNA genes. Isolate CadI showed 97.4% similarity to the 16S rRNA gene of a Flavobacterium strain, unlike CadII which showed 99.7% similarity to the 16S rRNA gene of a Sphingomonas paucimobilis. Both isolates rapidly metabolized cadusafos in MSMN and SEM within 48 h with concurrent population growth. This is the first report for the isolation and characterization of soil bacteria with the ability to degrade rapidly cadusafos and use it as a carbon source. Degradation of cadusafos by both isolates was accelerated when MSMN was supplemented with glucose. In contrast, addition of succinate in MSMN marginally reduced the degradation of cadusafos. Both isolates were also able to degrade completely ethoprophos, a nematicide chemical analog of cadusafos, but did not degrade the other organophosphorus nematicides tested such as isazofos and isofenphos. Inoculation of a soil freshly treated with cadusafos or ethoprophos (10 mg l(-1)) with high inoculum densities (4.3 x 10(8) cells g(-1)) of Sphingomonas paucimobilis resulted in the rapid degradation of both nematicides. These results indicate the potential of this bacterium to be used in the clean-up of contaminated pesticide waste in the environment.

31P MAS NMR: a useful tool for the evaluation of VX natural weathering in various urban matrixes.

The fate of chemical warfare agent VX (O-ethyl S-[2-(diisopropylamino)ethyl] methylphosphonothioate) in various urban matrixes was monitored utilizing 31P MAS NMR. Chosen matrixes represent buildings, roads, pavement, and earth found in urban environments. In view of the high toxicity of VX, solid state NMR afforded a fairly safe experimental mode, omitting any chance for evaporation. Moreover, due to the nondestructive nature of these experiments, measurements could be repeated over and over using the same samples. Degradation rates of VX were obtained and compared to provide a list of relative reactivity toward VX: concrete >> desert sand > beach sand > asphalt approximately to bitumen sheet. Chemical interactions between VX, its degradation products, and the matrixes were often expressed by widening of the peaks to the extent that mass balance could not be achieved. It is noteworthy that these experiments were usually carried out on crushed or milled specimens, allowing high reactivity and rapid reactions.

Enhanced microbial degradation of cadusafos in soils from potato monoculture: demonstration and characterization.

Rapid degradation of cadusafos was evident in soils collected from previously-treated field sites from a potato monoculture area in northern Greece. The slower degradation of cadusafos observed in corresponding antibiotic-treated soils as well as in soils from an adjacent previously-untreated field demonstrated the microbial involvement in the rapid degradation of cadusafos in the soils from the previously-treated sites. Application of the non-specific antibacterial antibiotic chloramphenicol or of the Gram+ bacteria-inhibiting antibiotics penicillin + lyncomycin + vancomycin significantly inhibited the rapid biodegradation of cadusafos suggesting that soil bacteria and probably Gram+ bacteria are mainly responsible for the rapid biodegradation of cadusafos in the specific soil. Further experiments showed that the bacterial population of the cadusafos-adapted soil was also able to rapidly degrade the chemically related nematicide ethoprophos but not fenamiphos and oxamyl. This is the first report of the occurrence of enhanced biodegradation of cadusafos in potato fields. In addition, the finding of cross-enhancement between cadusafos and ethoprophos significantly reduces the number of available chemicals which could be alternated to prevent the development of enhanced biodegradation and thus intensifies the problem in potato monoculture areas like the one in northern Greece.

Physicochemical and biological data for the development of predictive organophosphorus pesticide QSARs and PBPK/PD models for human risk assessment.

A search of the scientific literature was carried out for physiochemical and biological data [i.e., IC50, LD50, Kp (cm/h) for percutaneous absorption, skin/water and tissue/blood partition coefficients, inhibition ki values, and metabolic parameters such as Vmax and Km] on 31 organophosphorus pesticides (OPs) to support the development of predictive quantitative structure-activity relationship (QSAR) and physiologically based pharmacokinetic and pharmacodynamic (PBPK/PD) models for human risk assessment. Except for work on parathion, chlorpyrifos, and isofenphos, very few modeling data were found on the 31 OPs of interest. The available percutaneous absorption, partition coefficients and metabolic parameters were insufficient in number to develop predictive QSAR models. Metabolic kinetic parameters (Vmax, Km) varied according to enzyme source and the manner in which the enzymes were characterized. The metabolic activity of microsomes should be based on the kinetic activity of purified or cDNA-expressed cytochrome P450s (CYPs) and the specific content of each active CYP in tissue microsomes. Similar requirements are needed to assess the activity of tissue A- and B-esterases metabolizing OPs. A limited amount of acetylcholinesterase (AChE), butyrylcholinesterase (BChE), and carboxylesterase (CaE) inhibition and recovery data were found in the literature on the 31 OPs. A program is needed to require the development of physicochemical and biological data to support risk assessment methodologies involving QSAR and PBPK/PD models.

Pirimiphos-methyl and benalaxyl losses in surface runoff from plots cultivated with potatoes.

Abstract: Losses of pirimiphos-methyl and benalaxyl in runoff water from clay soil plots cultivated with potatoes and of differing soil surface slopes were determined over approximately 120 days (1 October 1999-28 January 2000). The plot slopes were 0, 1, 2.5 and 5%, and soil erosion increased with the slope from 610 to 1760kgha(-1). The runoff of surface water was between 3.1 and 16.6% of the rainfall. Surface runoff was highest for the fifth and seventh runoff events due to rainfall, 51 days and 72 days after the first pesticide application. The maximum concentrations of the two pesticides in runoff occurred in the plots with the greatest slope (5%) during the fifth runoff event, November 21, 1999 reaching 8.4 and 12.3 microg litre(-1) for pirimiphos-methyl and 17.8 and 20.2 microg litre(-1) for benalaxyl in tilled and untilled plots respectively. The cumulative losses of pirimiphos-methyl in surface runoff from tilled and untilled plots with a slope 5% were estimated at only 0.37 and 0.59% of the initial applied active ingredient, respectively, while for plots with a slope 0% the percentages were 0.013 and 0.018%. For benalaxyl the corresponding values from tilled and untilled plots were 1.69 and 1.76% (slope 5%), and 0.062 and 0.085 (slope 0%). Degradation of the pesticides in the topsoil was monitored from October 1999 and May 2000. Cultivation of potatoes decreased the half-life of the two pesticides compared to the untilled fields, for pirimiphos-methyl from 16.7 to 9.2 days and for benalaxyl from 26.7 to 12.6 days. The slope of soil surface and the different sorption capacities for the compounds are the main parameters which influenced the transportation of studied pesticides, pirimiphos-methyl and benalaxyl residues via surface water in soil-water systems.

Short-term assay for the identification of neurotoxic compounds and their liver derived stable metabolites.

The release of stable neurotoxic metabolites from liver after metabolic activation of xenobiotics can be investigated in cultures of primary rat hepatocytes as metabolic activation system and embryonic chicken brain cell cultures as target. It was shown that adult rat hepatocyte cell cultures are a reliable bioactivating system for xenobiotics such as cyclophosphamide and isophenphos resulting in the release of stable metabolites into the supernatant. The cultured embryonic chicken brain cells were able to discriminate between the toxicity of parent drugs and its metabolite(s) and between metabolites with an unspecific cytotoxic activity (cyclophosphamide) and metabolites with a high potential to damage specific nerve cell population(s) (isophenphos). The observed neurotoxicity of the isophenphos metabolites is not an acute effect but induced only after a prolonged exposure period. The present study provides evidence that the subsequent use of hepatocytes and brain cell cultures has the potential to be used as an in vitro screening system for the identification of specific and non-specific neurotoxic compounds. Solely stable metabolites are taken into account, since in the in vivo situation only stable metabolites have a change to reach the nervous system. Our data suggest that the present approach is able to discriminate (a) between cell-type and organ specific toxicity, (b) between neurotoxicity derived after long-term or acute exposure, and (c) between the neurotoxicity induced by the native test compound or stable metabolites.